The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.
The 16S V2-V3 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V2-V3 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.
Storage Conditions and Product Stability Norgen’s 16S V2-V3 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.
All kit components should remain stable for at least 1 year when stored at the specified storage conditions.
Step
Component
Cat. 70300 (24 preps)
Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)
MGX Master Mix
330 µL
1,320 µL
16S V2-V3 Primer Mix
70 µL
280 µL
Index PCR (PCR 2)
Indexing Master Mix
660 µL
2 x 1,320 µL
N7xx Index Primer
50 µL
50 µL
S5xx Index Primer
70 µL
70 µL
PCR Clean-Up
Resuspension Buffer
2 x 1,250 µL
2 x 5,000 µL
Nuclease-free water
1,250 µL
1 x 6,000 µL
Other Products
D3141 HiPure Stool DNA Kit
Product Info
Document
Product Info
Introduction
With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.
At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.
This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 50-100mg stool samples
Applications
PCR, Southern Blot, enzyme digestion and NGS, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Stool
Sample amount
50-100mg
Yield
3-15μg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
750μl
Binding yield of column
100μg
Principle
Stool sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. Humic acid, proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.
Advantages
High purity – unique adsorbent can completely remove inhibitory factors
High concentration – maximum extraction of total DNA from stool samples
High recovery – DNA can be recovered at the level of PG
Good repeatability – silica technology can obtain ideal results every time
Kit Contents
Contents
D314102
D314103
Purification Times
50 Preps
250 Preps
HiPure DNA Mini Columns II
50
250
2ml Collection Tubes
50
250
2ml Bead Tubes
50
250
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
1.8 ml
10 ml
Buffer SPL
40 ml
200 ml
Buffer PCI
40 ml
200 ml
Buffer AL
20 ml
80 ml
Buffer GW1
22 ml
88 ml
Buffer GW2
20 ml
2 x 50 ml
Buffer AE
15 ml
30 ml
Storage and Stability
Proteinase K and Buffer PCI should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Document
With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.
At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.
This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
Features
5’→3’ DNA polymerase activity
No detectable 3’→5′ exonuclease (proofreading) activity
Generates PCR products with 3′-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
For direct loading into electrophoresis analysis
DNA bands can be visualized directly under UV or 470 nm blue light illumination
Applications
Routine PCR
Colony PCR
High throughput PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
Protected from light and avoid multiple freeze/thaw cycles 4°C for 6 months -20°C for 24 months
Document
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
CP0101J Triple Wrapped Irradiated Plate-Nutrient Agar
Product Info
Document
Product Info
Introduction
Usages: For monitoring and detection of surface of equipment and personnel hygiene.
Advantage: 1.This series of products was filling at A level of environment, and final sterilization by Gamma ray irradiation, triple packaging insure sterility and long shelf life. 2.Each dish was marking label product name, batch number, expiration date .Information is available of traceability. 3.Inner additional desiccant, reducing formation of condensation water, while inner additional sterile paper and plastic bags for easier transfer and cultivation. 4.Triple packing dense bags to avoid the penetration of hydrogen peroxide; clean gas was filled as a buffer to reduce broken bags and dish in transit.. 5.This series of products is available to store at (2-25 ℃), shelf life of up to 6 months.
Storage: Store in a cool (2-25 ℃), dry place, away from bright light. Storage period of 6 months.
